Disruption of BG14 results in enhanced callose deposition in developing seeds and decreases seed longevity and seed dormancy in Arabidopsis.

Seed longevity is an important trait for agriculture and the conservation of genetic resources. ß-1,3-Glucanases were first recognized as pathogenesis-related proteins involved in plant defense, but their roles in seeds are largely unknown. Here, we report a glycosylphosphatidylinositol-anchored ß-1,3-glucanase, BG14, that degrades callose in seed embryos and functions in seed longevity and dormancy in Arabidopsis. The loss of function of BG14 significantly decreased seed longevity, whereas functional reversion (RE) and overexpression (OE) lines reversed and increased the impaired phenotype, respectively. The loss of function of BG14 enhanced callose deposition in the embryos of mature seeds, confirmed by quantitative determination and the decreased callose degrading ability in bg14. The drop-and-see (DANS) assay revealed that the fluorescence signal in bg14 was significantly lower than that observed in the other three genotypes. BG14 is located on the periphery of the cell wall and can completely merge with callose at the plasmodesmata of epidermal cells. BG14 was highly expressed in developing seeds and was induced by aging and abscisic acid (ABA). The loss of function of BG14 led to a variety of phenotypes related to ABA, including reduced seed dormancy and reduced responses to treatment with ABA or pacolblltrazol, whereas OE lines showed the opposite phenotype. The reduced ABA response is because of the decreased level of ABA and the lowered expression of ABA synthesis genes in bg14. Taken together, this study demonstrated that BG14 is a bona fide BG that mediates callose degradation in the plasmodesmata of embryo cells, transcriptionally influences ABA synthesis genes in developing seeds, and positively affects seed longevity and dormancy in Arabidopsis.

Receptor-like kinase HAESA-like 1 positively regulates seed longevity in Arabidopsis.

MAIN CONCLUSION: Based on the phenotypic, physiological and transcriptomic analysis, receptor-like kinase HAESA-like 1 was demonstrated to positively affect seed longevity in Arabidopsis. Seed longevity is very important for both genetic resource conservation and crop production. Receptor-like kinases (RLKs) are widely involved in plant growth, development and stress responses. However, the role of most RLKs, especially in seed longevity, is largely unknown. In this study, we report that Arabidopsis HAESA-like 1 (AtHSL1) positively regulated seed longevity. Disruption of HSL1 significantly decreased the germination rate to 50% at 7 days after cold stratification (DAC), compared with that of the wild type (93.5% at 7 DAC), after accelerated aging treatment. Expression of the HSL1 gene in hsl1 basically restored the defective phenotype (86.3%), while HSL1-overexpressing lines (98.3%) displayed slower accelerated aging than WT (93.5%). GUS staining revealed HSL1 was highly expressed universally, especially in young seedlings, mature seeds and embryos of imbibed seeds, and its expression could be induced by accelerated aging. No difference in the dyeing color and area of mucilage were identified between WT and hsl1. The soluble pectin content also was not different, while the adherent pectin content was significantly increased in hsl1. Global transcriptomics revealed that disruption of HSL1 mainly downregulated genes involved in trehalose synthesis, nucleotide sugar metabolism and protection and repair mechanisms. Therefore, an increase in adherent pectin content and downregulation of genes involved in trehalose synthesis may be the main reasons for decreasing seed longevity owing to disruption of HSL1 in Arabidopsis. Our work provides valuable information for understanding the function and mechanism of a receptor-like kinase, AtHSL1, in seed longevity.

C/EBPß enhances immunosuppression activity of myeloid-derived suppressor cells by a P300-mediated acetylation modification.

OBJECTIVE: Myeloid-derived suppressor cells (MDSCs) are a major immunosuppressive population in the tumor microenvironment,inhibiting anti-tumor immune response and exerting pro-tumorigenic effect. CCAAT/enhancer-binding protein beta (C/EBPß), a key transcription factor indispensable for myelopoiesis, plays a fundamental role in regulating expansion and activation of MDSCs. Lysine acetylation can regulate functions of transcription factors. However, the role of C/EBPß acetylation modification in MDSCs has not been reported. MATERIALS AND METHODS: MDSCs derived from the spleens of tumor-bearing mice (TB-SP-MDSCs) were isolated by immunomagnetic beads. Bone marrow derived MDSCs were induced by IL-6 and GM-CSF. Western-blot was used to detect the expression of P300 and co-immunoprecipitation (CO-IP) was used to detect the C/EBPß acetylation in MDSCs. Inhibitor C646 was used to specificly inhibit P300 activity. RESULTS: In this study, we found that C/EBPß was acetylated by acetyltransferase P300 in MDSCs. A P300-mediated C/EBPß acetylation enhanced C/EBPß transactivation activity on arginase 1 (Arg-1) gene promoter. Inhibition of P300 activity downregulated the inhibitory effects of MDSCs in vitro and attenuated pro-tumorigenic effects of MDSCs in vivo. Additionally, IL-6 from tumor microenvironment could upregulate the expression of P300 and enhance C/EBPß acetylation in MDSCs. CONCLUSION: In general, a P300-mediated C/EBPß acetylation enhanced C/EBPß transactivation activity on Arg-1 promoter, thus promoting immunosuppressive function of MDSCs. In view of the critical role of P300 in regulating MDSCs, P300 might be a potential target of anti-tumor immunotherapy.

Thiol-acrylate side-chain liquid crystal elastomers.

The Michael addition 'click' chemistry was used to graft acrylate-terminated mesogenic groups onto the polysiloxane backbone polymer chain with thiol functional groups, with a constant 15% fraction of diacrylate reacting monomers as crosslinkers. Three different types of mesogens were used, and also their 50 : 50 mixtures, and in all cases we have obtained the smectic-A phase of the resulting liquid crystalline elastomer. Using X-ray diffraction, calorimetry and dynamic mechanical analysis, we investigated the relationship between the molecular structure of mesogenic side groups and the structure and properties of the elastomers. The shape-memory of smectic elastomers was verified. The unusual features were the semi-crystalline nature of elastomers with non-polar mesogens and the clear role of side-by-side rod dimerization of polar mesogens leading to a higher smectic layer spacing. We investigated the evolution of the smectic alignment on uniaxial stretching along the layer normal and identified two distinct ways in which the elastomer responds: the coarsened Helfrich-Hurault zig-zag layer texture and the large-scale stripe domains of uniform layer rotation in the systems with lower order parameter and the associated layer constraints.

Synthesis and Anti-Inflammatory Activity of 1-Methylhydantoin Cinnamoyl Imides.

In this study, 1-methylhydantoin cinnamic imides were synthesized from 1-methylhydantoin and trans-cinnamic acid, and their anti-inflammatory activity was investigated. The anti-inflammatory activity in vitro was evaluated by measuring the contents of NO, TNF-α and IL-1ß in the supernatant of RAW264.7 cells stimulated by LPS. The cytotoxicity of 1-methylhydantoin cinnamoyl imides on RAW264.7 cells was detected using the CCK-8 method. The results showed that compounds 2 and 4 can significantly inhibit the release of NO and reduce the secretion of TNF-α and IL-1ß. Compound 3 inhibited the production of TNF-α. The inhibition rate of COX was evaluated in vitro. The in vivo anti-inflammatory activities of the five compounds were evaluated by establishing an animal model of xylene ear swelling. The results showed that 1-methylhydantoin cinnamic imides could alleviate xylene-induced ear edema in mice in a dose-dependent manner. Among them, the effect of compound 5 was the most significant. Under the action of high dosage, its ear swelling inhibition rate was as high as 52.08%.

Increased GITRL Impairs the Function of Myeloid-Derived Suppressor Cells and Exacerbates Primary Sjögren Syndrome.

Although the expansion of myeloid-derived suppressor cells (MDSCs) has been reported in autoimmune disorders, it is largely unclear how MDSCs contribute to the development of primary Sjögren syndrome (pSS). In this study, we found significantly increased MDSCs with gradually diminished suppressive capacity during disease development in mice with experimental Sjögren syndrome (ESS). The ligand for glucocorticoid-induced TNFR family-related protein (GITRL) was increased along ESS progression, whereas the increased GITRL was found to attenuate the immunosuppressive function of MDSCs. Moreover, blocking GITR signal in MDSCs significantly restored their immunosuppressive function and alleviated ESS progression in mice. In pSS patients, expanded MDSCs were found to express low levels of arginase. Significantly increased serum GITRL levels were closely correlated with patients with higher Sjögren syndrome disease activity index. Furthermore, treatment with recombinant GITRL markedly reduced the immunosuppressive function of human MDSCs. Together, our studies have demonstrated a critical role of GITRL in modulating the suppressive function of MDSCs, which may facilitate the validation of GITRL as a therapeutic target for the treatment of pSS.

Crystal Engineering Construction of Caffeic Acid Derivatives with Potential Applications in Pharmaceuticals and Degradable Polymeric Materials.

Natural products are precious feedstock in drug discovery and sustainable materials. This work using crystal engineering strategy, visible light, and solvent-free cycloaddition successfully constructed two caffeic acid derivatives, rel-(1R,2R,3S,4S)-2,4-bis(3,4-dihydroxyphenyl)cyclobutane-1,3-dicarboxylate and rel-(1R,2R,3S,4S)-2,4-bis(3,4-dihydroxyphenyl)cyclobutane-1,3-dicarboxylic acid. Because of the multiple stereocenters, it is challenging to prepare those compounds using traditional organic synthesis methods. The crystal engineering Hirshfeld surface analysis and 2D intermolecular interaction fingerprints were applied to synthetic route design. The light resources used in this work was visible LED or free, clean, and renewable sunlight. The evidence suggested that pure stereoisomer was obtained demonstrating the stereospecificity and efficiency of the topochemical cycloaddition reaction. The derivatives exhibited free radical scavenging and antioxidant biological activities, as well as the potential inhibitory activity of fatty acid binding proteins. One of the derivatives is the precursor of the natural product Shimobashiric acid C which paves the way for the total synthesis and further study of Shimobashiric acid C. In addition, the derivatives possess photodegradability at a specific wavelength, which is very attractive for "green" degradable polymeric materials.

Rana chensinensis Ovum Oil Based on CO2 Supercritical Fluid Extraction: Response Surface Methodology Optimization and Unsaturated Fatty Acid Ingredient Analysis.

Rana chensinensis ovum oil (RCOO) is an emerging source of unsaturated fatty acids (UFAs), but it is lacking in green and efficient extraction methods. In this work, using the response surface strategy, we developed a green and efficient CO2 supercritical fluid extraction (CO2-SFE) technology for RCOO. The response surface methodology (RSM), based on the Box-Behnken Design (BBD), was used to investigate the influence of four independent factors (pressure, flow, temperature, and time) on the yield of RCOO in the CO2-SFE process, and UPLC-ESI-Q-TOP-MS and HPLC were used to identify and analyze the principal UFA components of RCOO. According to the BBD response surface model, the optimal CO2-SFE condition of RCOO was pressure 29 MPa, flow 82 L/h, temperature 50 °C, and time 132 min, and the corresponding predicted optimal yield was 13.61%. The actual optimal yield obtained from the model verification was 13.29 ± 0.37%, and the average error with the predicted value was 0.38 ± 0.27%. The six principal UFAs identified in RCOO included eicosapentaenoic acid (EPA), α-linolenic acid (ALA), docosahexaenoic acid (DHA), arachidonic acid (ARA), linoleic acid (LA), and oleic acid (OA), which were important biologically active ingredients in RCOO. Pearson correlation analysis showed that the yield of these UFAs was closely related to the yield of RCOO (the correlation coefficients were greater than 0.9). Therefore, under optimal conditions, the yield of RCOO and principal UFAs always reached the optimal value at the same time. Based on the above results, this work realized the optimization of CO2-SFE green extraction process and the confirmation of principal bioactive ingredients of the extract, which laid a foundation for the green production of RCOO.

Protein-Based Fingerprint Analysis for the Identification of Ranae Oviductus Using RP-HPLC.

This work demonstrated a method combining reversed-phase high-performance liquid chromatography (RP-HPLC) with chemometrics analysis to identify the authenticity of Ranae Oviductus. The fingerprint chromatograms of the Ranae Oviductus protein were established through an Agilent Zorbax 300SB-C8 column and diode array detection at 215 nm, using 0.085% TFA (v/v) in acetonitrile (A) and 0.1% TFA in ultrapure water (B) as mobile phase. The similarity was in the range of 0.779-0.980. The fingerprint chromatogram of Ranae Oviductus showed a significant difference with counterfeit products. Hierarchical clustering analysis (HCA) and principal component analysis (PCA) successfully identified Ranae Oviductus from the samples. These results indicated that the method established in this work was reliable.

Antitussive and Anti-inflammatory Dual-active Agents Developed from Natural Product Lead Compound 1-Methylhydantoin.

Natural products play an important role in drug discovery. This work employed a natural product 1-methylhydantoin as the lead compound to develop novel dual-active drugs. 1-Methylhydantoin was isolated from Oviductus Ranae, which is a traditional Chinese medicine that has been used for tussive and inflammation treatment for a long time. An in silico study screened the more active 1-methylhydantoin derivatives. Antitussive assessment indicated that the newly synthesized agent had similar bioactivity with the natural product. An anti-inflammatory model used xylene induced ear edema model. At the same dosage (100 mg/Kg), the newly prepared agent had an inhibition rate 53.18% which was much higher than that of the lead compound (22.69%). The results might be ascribed to the cyclooxygenases-1 (COX-1) and cyclooxygenases-2 (COX-2) selectivity, and the fitness of the compound, and the binding pocket. The anti-particulate matter (PM 2.5) acute pneumonia was evaluated through an in vivo model constructed by nasal instillation with PM 2.5 suspension. The results of the above models suggested that this novel agent had remarkable antitussive, anti-inflammatory, and anti-PM 2.5 acute pneumonia activities.

Modeling and Performance of the IEEE 802.11p Broadcasting for Intra-Platoon Communication.

Road capacity, traffic safety, and energy efficiency can be extremely improved by forming platoons with a small intra-vehicle spacing. Automated controllers obtain vehicle speed, acceleration, and position through vehicular ad hoc networks (VANETs), which allows the performance of platoon communication to make a significant impact on the stability of the platoon. To the best of our knowledge, there is not much research relating to packet delay and packet dropping rate of platoon communication based on the IEEE 802.11p broadcasting. In this paper, we introduce platoon structure model, vehicle control model, and communication model for a single platoon scenario. By utilizing Markov process and M/G/1/K queuing theory, we put forward an analytical model to assess the property of intra-vehicle communication. The analytical model is validated by simulations and the influence of communication parameters on intra-vehicle communication performance are discussed. In addition, the experimental results demonstrate that the IEEE 802.11p-based intra-vehicle communication guarantee the stability of platoon.

Conjugates of 18ß-glycyrrhetinic acid derivatives with 3-(1H-benzo[d]imidazol-2-yl)propanoic acid as Pin1 inhibitors displaying anti-prostate cancer ability.

Twenty-six conjugates of 18ß-glycyrrhetinic acid derivatives with 3-(1H-benzo[d]imidazol-2-yl)propanoic acid were designed and synthesized as Pin1 inhibitors. Most of these semi-synthetic compounds showed improved Pin1 inhibitory activity and anti-proliferative effects against prostate cancer cells as compared to 3-(1H-benzo[d]imidazol-2-yl)propanoic acid and GA. Compounds 10a and 12i were the most potent to inhibit growth of prostate cancer PC-3 with GI50 values of 7.80µM and 3.52µM, respectively. The enzyme inhibition ratio of nine compounds at 10µM was over 90%. Structure-activity relationships indicated that both appropriate structure at ring C of GA and suitable length of linker between GA skeleton and benzimidazole moiety had significant impact on improving activity. Western blot assay revealed that 10a decreased the level of cell cycle regulating protein cyclin D1. Thus, these compounds might represent a novel anti-proliferative agent working through Pin1 inhibition.

Synthesis and biological evaluation of quinoline derivatives as potential anti-prostate cancer agents and Pim-1 kinase inhibitors.

In this work, a series of quinoline derivatives were designed and synthesized as antitumor agents. Most quinolines showed potent anti-proliferative activity against human prostatic cancer PC-3 cell line. Among which, 9d, 9f and 9g were the most effective compounds with GI50 values of 2.60, 2.81 and 1.29 µM, respectively. Structure-activity relationship analysis indicated that the secondary amine linked quinoline and pyridine ring played an important role in the anti-proliferative effects. Mechanistic studies revealed that 9g was a potential Pim-1 kinase inhibitor with abilities of cell cycle arrest and apoptosis induction. Considering of the increased activity of Pim-1 in prostate cancer, such compounds have potential to be developed as anti-prostate cancer agents.

ß-Glucan enhances antitumor immune responses by regulating differentiation and function of monocytic myeloid-derived suppressor cells.

Myeloid-derived suppressor cells (MDSCs) accumulate in tumor-bearing hosts and play a major role in tumor-induced immunosuppression, which hampers effective immuno-therapeutic approaches. ß-Glucans have been reported to function as potent immuno-modulators to stimulate innate and adaptive immune responses, which contributes to their antitumor property. Here, we investigated the effect of particulate ß-glucans on MDSCs and found that ß-glucan treatment could promote the differentiation of M-MDSCs (monocytic MDSCs) into a more mature CD11c(+) F4/80(+) Ly6C(low) population via dectin-1 pathway in vitro, which is NF-κB dependent, and the suppressive function of M-MDSCs was significantly decreased. Treatment of orally administered yeast-derived particulate ß-glucan drastically downregulated MDSCs but increased the infiltrated DCs and macrophages in tumor-bearing mice, thus eliciting CTL and Th1 responses, inhibiting the suppressive activity of regulatory T cells, thereby leading to the delayed tumor progression. We show here for the first time that ß-glucans induce the differentiation of MDSCs and inhibit the regulatory function of MDSCs, therefore revealing a novel mechanism for ß-glucans in immunotherapy and suggesting their potential clinical benefit.

[Optimization of formulas of Quban gel by uniform design].

OBJECTIVE: To optimize formulas of Quban gel. METHOD: The U6 (6(2) x 3) uniform design was adopted to optimize gel formulas, with rheological parameters, such as viscosity and yield value in room temperature, viscosity and yield value in average temperature of skin, thixlotropy. RESULT: The optimum proportion of matrix was made of 1.0 g carbomer 940, 5 mL glycerin and pH value 5-6. CONCLUSION: The regression model for gel matrix quality and gel rheological parameters was established to directly reflect the impacting effect of various factors, and provide certain preference basis for the screening of gel matrix formulas. Quban gel prepared by the method was evenly distributed, moderately viscous and highly thixotropic

Mechanism of Pressure-Sensitive Adhesion in Nematic Elastomers.

Nematic liquid crystal elastomers (LCEs) have anomalously high vibration damping, and it has been assumed that this is the cause of their anomalously high-pressure-sensitive adhesion (PSA). Here, we investigate the mechanism behind this enhanced PSA by first preparing thin adhesive tapes with LCE of varying cross-linking densities, characterizing their material and surface properties, and then studying the adhesion characteristics with a standard set of 90° peel, lap shear, and probe tack tests. The study confirms that the enhanced PSA is only present in (and due to) the nematic phase of the elastomer, and the strength of bonding takes over 24 h to fully reach its maximum value. Such a long saturation time is caused by the slow relaxation of local stress and director orientation in the nematic domains after pressing against the surface. We confirm this mechanism by showing that freshly pressed and annealed tape reaches the same maximum bonding strength on cooling, when the returning nematic order is forming in its optimal configuration in the pressed film.

Controlling factors and health risks of groundwater chemistry in a typical alpine watershed based on machine learning methods.

Groundwater is a key water resource in alpine watersheds, but its quality is deteriorating due to human activities. The Golmud River watershed is a representative alpine watershed in Northwest China, and it was chosen to explore groundwater chemistry, associated controlling factors, source contributions, and potential health risks. The analysis includes the use of a self-organizing map (SOM), positive matrix factorization (PMF), ionic ratios, and a Monte Carlo simulation. The content of total dissolved solids in phreatic water was higher in the dry season and increased from the mountainous zone to the fine-soil plain-overflowing zone. Additionally, the water type varied from HCO3- to Cl- types whereas confined groundwater was chemically stable and of a HCO3- type. The SOM results showed a visual correlation between the ions in groundwater. The combination of SOM, PMF, and ionic ratios identified water-rock action as a dominant factor of groundwater chemistry. It was also found that Clusters I and III were mainly influenced by silicate weathering (a total contribution of 38.4 %), whereas evaporation was dominant in Cluster VI (a contribution of 32.5 %). Anthropogenic pollution was mainly associated with clusters V and IV and was related to industrial and agricultural activities during the snowmelt and wet seasons, and fluorine deposition formed by residential coal heating during the dry season (contributions of 1.4 % and 23.8 % in Clusters V and IV, respectively). The sudden increases in B3+ and Li+ in Cluster II were due to inputs from small tributaries (a contribution of 3.9 %). The probabilistic health risk assessment showed that fluoride posed a greater non-carcinogenic risk to human health than Sr2+, B3+, and NO3-, and its potential threat to children was more significant during the dry season than in other seasons. It is necessary for local governments to establish urgent fluoride emission control policies within the Golmud River watershed.

Momentum transfer on impact damping by liquid crystalline elastomers.

The effect of elastomeric damping pads, softening the collision of hard objects, is investigated comparing the reference silicone elastomer and the polydomain nematic liquid crystalline elastomer, which has a far superior internal dissipation mechanism. We specifically focus not just on the energy dissipation, but also on the momentum conservation and transfer during the collision, because the latter determines the force exerted on the target and/or the impactor-and it is the force that does the damage during the short time of an impact, while the energy might be dissipated on a much longer time scale. To better assess the momentum transfer, we compare the collision with a very heavy object and the collision with a comparable mass, when some of the impact momentum is retained in the target receding away from the collision. We also propose a method to estimate the optimal thickness of an elastomer damping pad for minimising the energy in impactor rebound. It has been found that thicker pads introduce a large elastic rebound and the optimal thickness is therefore the thinnest possible pad that does not suffer from mechanical failure. We find good agreement between our estimate of the minimal thickness of the elastomer before the puncture through occurs and the experimental observations.

Granulocytic Myeloid-Derived Suppressor Cell Exosomal Prostaglandin E2 Ameliorates Collagen-Induced Arthritis by Enhancing IL-10+ B Cells.

The results of recent studies have shown that granulocytic-myeloid derived suppressor cells (G-MDSCs) can secrete exosomes that transport various biologically active molecules with regulatory effects on immune cells. However, their roles in autoimmune diseases such as rheumatoid arthritis remain to be further elucidated. In the present study, we investigated the influence of exosomes from G-MDSCs on the humoral immune response in murine collagen-induced arthritis (CIA). G-MDSCs exosomes-treated mice showed lower arthritis index values and decreased inflammatory cell infiltration. Treatment with G-MDSCs exosomes promoted splenic B cells to secrete IL-10 both in vivo and in vitro. In addition, a decrease in the proportion of plasma cells and follicular helper T cells was observed in drainage lymph nodes from G-MDSCs exosomes-treated mice. Moreover, lower serum levels of IgG were detected in G-MDSCs exosomes-treated mice, indicating an alteration of the humoral environment. Mechanistic studies showed that exosomal prostaglandin E2 (PGE2) produced by G-MDSCs upregulated the phosphorylation levels of GSK-3ß and CREB, which play a key role in the production of IL-10+ B cells. Taken together, our findings demonstrated that G-MDSC exosomal PGE2 attenuates CIA in mice by promoting the generation of IL-10+ Breg cells.

Mesenchymal Stem Cell Enhances the Function of MDSCs in Experimental Sjögren Syndrome.

Primary Sjögren's syndrome (pSS) is a progressive systemic autoimmune disease characterized by lymphocytic infiltrates in exocrine glands, leading to the injury of salivary and lachrymal glands. Mesenchymal stem cells (MSCs) have been demonstrated to exert great potential in the treatment of various autoimmune diseases. Although MSCs have provide an effective therapeutic approach for SS treatment, the underlying mechanisms are still elusive. Our previous study has shown the reduced suppressive capacity of myeloid-derived suppressor cells (MDSCs) advanced the progression of experimental Sjögren's syndrome (ESS). In this study, we found that BM-MSCs significantly enhanced the suppressive function of MDSCs with high levels of Arginase and NO, decreased the levels of CD40, CD80, CD86, and MHC-II expression on MDSCs, thus attenuating the disease progression in ESS mice. Furthermore, the enhanced suppressive function of MDSCs was mediated by BM-MSC-secreted TGF-ß, and the therapeutic effect of BM-MSCs in inhibiting ESS was almost abolished after silencing TGF-ß in BM-MSCs. Taken together, our results demonstrated that BM-MSCs alleviated the ESS progression by up-regulating the immunosuppressive effect of MDSCs through TGF-ß/Smad pathway, offering a novel mechanism for MSCs in the treatment of pSS.